Abstract
An efficient protocol for direct shoot organogenesis in Bixa orellana, known as achiote or annatto or Latkhan (India), has been developed. Using nodal shoot-tip explants, significant organogenetic responses, mean shoot number and shoot elongation were observed when these were incubated on Murashige and Skoog (MS) medium containing 6.66 μM 6-benzyladenine (BA) and 4.9 μM indole-3-butyric acid (IBA), and supplemented with either 200–1,500 μM putrescine or 40 μM silver nitrate (AgNO3). Putrescine at 800 and 1,000 μM promoted the highest mean shoot length and mean shoot number per explant, respectively. Moreover, various concentrations of putrescine induced callus development. Incorporation of a polyamine biosynthesis inhibitor Difluro-Methyl Ornithine (DFMO) inhibited in vitro shoot multiplication and also altered the endogenous polyamine pool of B. orellana shoots. The field survival of in vitro-derived plants of putrescine and AgNO3 treatments was 70%. This protocol can be used for improving the in vitro regeneration of B. orellana for transformation studies.
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References
Aparnathi K, Lata R, Sharma R (1990) Annatto (Bixa orellana L.): its cultivation preparation and usage. Int J Trop Agric 8:80–88
Bais HP, Ravishankar GA (2002) Role of polyamines in the ontogeny of plants and their biotechnological applications. Plant Cell Tiss Organ Cult 69:1–34
Bais HP, Sudha GS, Ravishankar GA (2000) Putrescine and silver nitrate influences shoot multiplication, in vitro flowering and endogenous titers of polyamines in Chichorium intybus L. cv. Lucknow Local. J Plant Growth Regul 19:238–248
Beyer EM (1976) A potent inhibitor of ethylene action in plants. Plant Physiol 58:268–271
D’Souza MC, Sharon M (2001) In vitro clonal propagation of annatto (Bixa orellana L.). In Vitro Cell Dev Biol Plant 37:168–172
De Paiva Neto VB, Ribeiro da Mota T, Otoni WC (2003) Direct organogenesis from hypocotyl-derived explants of annatto (Bixa orellana). Plant Cell Tiss Organ Cult 75:159–167
Evans PT, Malmberg RL (1989) Do polyamines have a role in plant development? Annu Rev Plant Physiol Mol Biol 40:235–269
Fierer RP, Mignon G, Litvay JD (1984) Arginine decarboxylase and polyamine required for embryogenesis in wild carrot. Science 223:1433–1435
Flores HE, Galston AW (1982) Analysis of polyamines in higher plants by HPLC. Plant Physiol 69:701–706
Galston AW, Kaur-Sawhrey R (1990) Polyamines in plant physiology. Plant Physiol 94:406–410
Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158
Gerats AGM, Keye C, Collins C, Malmberg RL (1988) Polyamine level in Petunia genotypes with normal and abnormal floral morphologies. Plant Physiol 86:390–393
Giridhar P, Reddy SM (1998) The effect of some polyamine biosynthesis inhibitors on mycelial growth and mycotoxin production by Aspergillus terreus. Nat Acad Sci Lett India 21:5–8
Giridhar P, Reddy SM, Rajam MV (1997) Control of Penicillium citrinum growth and citrinin production by some polyamine biosynthesis inhibitors. Ind Phytopathol Soc 50(1):33–36
Giridhar P, Rajam MV, Reddy SM (1999) Effect of polyamine biosynthesis inhibitors on aflatoxin B1 and ochratoxin a production. Adv Plant Sci 12(1):41–44
Giridhar P, Obul Reddy B, Ravishankar GA (2001) Silver nitrate influences in vitro shoot multiplication and root formation in Vanilla planifolia. Curr Sci 81(9):101–103
Harborne JB (1975) Flavanoid bisulphates and their co-occurrence with ellagic acid in the Bixaceae, Frankineaceae and related families. Phytochem 14:1331–1337
Jarvis BC, Yasmin S, Coleman MT (1985) RNA and protein metabolism during adventitious root formation in stem cuttings of P. mungo cultivar berkin. Physiol Plant 64:53–59
Kallio A, McCann PP (1981) Difluromethylornithine irreversibly inactivates ornithine decarboxylase of Pseudomonas areuginosa, but does not inhibit the enzymes of Escherichia coli. Biochem J 200:69–75
Kumar V, Giridhar P, Ravishankar GA (2009) AgNO3—a potential regulator of ethylene activity and plant growth modulator. Electron J Biotechnol 12(2). Issue of April 15
Leonardo LCD, Santa-Catarina C, Ribeiro DM, Barros RS, Floh EIS, Wagner CO (2009) Ethylene and polyamine production patterns during in vitro shoot organogenesis of two passion fruit species as affected by polyamines and their inhibitor. Plant Cell Tiss Organ Cult 101:163–170
Meena K, Abraham CJ, Mani B, Thomas TD (2010) Adventitious shoot induction from cultured internodal explants of Malaxis acuminata D. Don, a valuable terrestrial medicinal orchid. Plant Cell Tiss Organ Cult 99:199–208
Murashige M, Skoog T (1962) A revised medium for rapid growth and bioassay of tobacco tissue cultures. Physiol Plant 15:473–497
Parimalan R, Giridhar P, Gururaj HB, Ravishankar GA (2007a) Organogenesis from cotyledon and hypocotyl-derived explants of japhara (Bixa orellana L.). Acta Bot Croat 66:153–160
Parimalan R, Giridhar P, Rajasekaran T, Ravishankar GA (2007b) Annatto fruit pericarp: newer source as a potential fuel. Energy Fuels 21:1181–1182
Parimalan R, Giridhar P, Ravishankar GA (2008) Mass multiplication of Bixa orellana L. through tissue culture for commercial propagation. Ind Crops Prod 28:122–127
Parimalan R, Giridhar P, Gururaj HB, Ravishankar GA (2009) Micropropagation of Bixa orellana using phytohormones and triacontanol. Biol Plantarum 53(2):347–350
Petri C, Nuria Alburquerque N, Pérez-Tornero O, Burgos L (2005) Auxin pulses and a synergistic interaction between polyamines and ethylene inhibitors improve adventitious regeneration from apricot leaves and Agrobacterium-mediated transformation of leaf tissues. Plant Cell Tiss Org Cult 82:105–111
Pradeep CD, Harding RM, Taylor M, Tyagi AP, Becker DK (2009) Somatic embryogenesis, organogenesis and plant regeneration in taro (Colocasia esculenta var. esculenta). Plant Cell Tiss Organ Cult 99:61–71
Pua EC, Sim GE, Chi GL, Kong LF (1996) Synergistic effect of ethylene inhibitors and putrescine on shoot regeneration from hypocotyl explants of Chinese radish (Raphanus sativus L. var. longipinnatus Bailey) in vitro. Plant Cell Rep 15:685–690
Ramamurthy N, Savithramma R, Swamy PM (1999) Multiple shoot induction and regeneration of Japhara (Bixa orellana L.) through axillary bud derived callus cultures. J Plant Biol 26:231–235
Smith TA (1990) Plant polyamines metabolism and function. In: Flores HE, Arteea RN, Shannon JC (eds) Polyamines and ethylene: biochemistry, physiology and interaction. American Society of Plant Physiologists, Maryland, pp 1–23
Sridevi V, Giridhar P, Ravishankar GA (2009) Endogenous polyamine profiles in different tissues of Coffea sp., and their levels during the ontogeny of fruits. Acta Physiol Plantarum 31:757–764
Sridevi V, Giridhar P, Simmi PS, Ravishankar GA (2010) Direct shoot organogenesis on hypocotyls explants with collar region from in vitro seedlings of Coffea canephora Pierre ex. Frohner cv. CxR and Agrobacterium tumefaciens- mediated transformation. Plant Cell Tiss Organ Cult 101:339–347
Srivastava A, Shukla YN, Jain S, Kumar S (1999) Chemistry, pharmacology and uses of Bixa orellana: a review. J Med Aromat Plant Sci 21:1145–1154
Steel RGD, Torrie JH (1980) Principles, and procedures of statistics. McGraw Hill, New York
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This work was supported by a research grant from Department of Science and Technology, Government of India. Author R.P. is grateful to CSIR for a research fellowship.
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Parimalan, R., Giridhar, P. & Ravishankar, G.A. Enhanced shoot organogenesis in Bixa orellana L. in the presence of putrescine and silver nitrate. Plant Cell Tiss Organ Cult 105, 285–290 (2011). https://doi.org/10.1007/s11240-010-9865-7
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DOI: https://doi.org/10.1007/s11240-010-9865-7